【摘要】 目的:比较转染Survivin、bcl -2反义寡核苷酸(ASODN)对胆囊癌细胞系 GBC SD的凋亡促进作用。方法:设计并合成特异性靶向 Survivin及 bcl- 2 的 ASODN。胆囊癌细胞分为 5 组:空白对照组、Survivin ASODN转染组、bcl- 2 ASODN转染组、Survivin SODN转染组和bcl -2 SODN转染组,转染24h后,采用逆转录 聚合酶链反应(RT PCR)检测Survivin基因、bcl- 2基因表达的变化,流式细胞仪技术检测细胞凋亡率,四甲基偶氮唑蓝(MTT)法检测 ASODN对细胞的生长抑制率。结果:转染 Survivin及bcl 2 ASODN后,胆囊癌细胞内Survivin基因表达下调 47.8%,bcl- 2 基因表达下调 52.4%;细胞凋亡率分别为(11.4±3. 9)%、(7. 3±3. 2)%,与对照组差异有统计学意义(P< 0. 01)。而转染 SurvivinASODN组及bc1 2ASODN组相比差异也有显著性(P<0.05)。两转染组相对于空白对照组的生长抑制率分别为54.3%、41.6%。结论:Survivin和bcl 2反义寡核苷酸均可有效的抑制Survivin及bc1- 2基因的表达,从而诱导胆囊癌细胞的凋亡,两者相比,Survivin反义寡核苷酸可更为有效地诱导胆囊癌细胞的凋亡。更多还原

【Abstract】 Objective:To investigate the apoptosis induced by antisense oligonucleotide targeting Survivin and bc1-2 on gallbladder cancer line GBC-SD in vitro.Methods:ASODN targeting Survivin and bc1-2 was designed and constructed.Cultrued cells were divided into 5 groups:control group,Survivin ASODN group,bcl-2 ASODN group,Survivin SODN group,bcl-2 SODN group.After transfected for 24 h,the cultured cells were harvested to carry out the next tests.The expression of Survivin and bc1-2 was detected by RT-PCR;Apoptosis index(AI) was examined by flow cytometry;Inhibitory rate(IR)by ASODN was determined by the colorimetri MTT cell viability and proliferation assay.Results:The expression of Survivin and bc1-2 were decreased 47.83% and 52.35% respectively.The AI in Survivin ASODN group and bc1-2 ASODN group was 11.38%± 3.91% and 7.26%±3.15% respectively and significantly higher than the control group(P<0.01),and that in the Survivin ASODN group was significantly higher than the bc1-2 ASODN group,(P<0.05).The IR was 54.3% and 41.6% respectively.Conclusion:The expression of Survivin and bcl-2 may be decreased in GBC-SD cells after Survivin and bc1-2 ASODN transfection,and the apoptosis was induced.Comparing the two ASODNs,Survivin ASODN was more effectively.更多还原